Department of Gastrointestinal Surgery, The First Hospital of China Medical University, Shenyang, China.
PeerJ. 2022 Mar 23;10:e13172. doi: 10.7717/peerj.13172. eCollection 2022.
The Numb protein plays a vital role in tumor development. The main aim of this study was to identify ATP11A, which is associated with the biological behavior of pancreatic cancer, and elucidate its relationship with Numb and the underlying mechanism behind this relationship.
First, data retrieved from The Cancer Genome Atlas (TCGA) and Genotype-Tissue Expression (GTE) databases was used to investigate the expression of ATP11A mRNA and its relationship with Numb mRNA in pancreatic cancer. Western blot assays on 31 pairs of pancreatic cancer tissues and paracancerous tissues, and immunohistochemical assays on 81 pancreatic cancer specimens were performed in order to verify the expression of ATP11A in pancreatic cancer at the protein level. Next, ATP11A was overexpressed or knocked down to observe its effects on the invasion and migration ability of pancreatic cancer cells and the changes of downstream proteins. Rescue assays were conducted to determine the mechanism through which ATP11A affects Numb, ZEB1, Snail2 and other proteins. Furthermore, immunoprecipitation assays were performed to explore the interaction between ATP11A and Numb. Finally, pancreatic cancer cells were stimulated with TGFB1 and ATP11A expression was examined to explore whether the effect of ATP11A on EMT was TGFB dependent.
At the mRNA level, the expression of ATP11A in pancreatic cancer tissues was significantly higher than in normal pancreatic tissues ( < 0.001). ATP11A expression was also highly correlated with Numb expression (R = 0.676). At the protein level, ATP11A expression in pancreatic cancer tissues was significantly higher than that in paracancerous tissues ( = 0.0009), and high ATP11A expression was also correlated with a worse prognosis. Moreover, our results showed that ATP11A can promote the invasion and migration of pancreatic cancer cells. Additionally, ATP11A could positively regulate the expression of Numb PRR, Snail2 and ZEB1 proteins. The rescue experiment results showed that the enhancement effect of ATP11A on ZEB1/Snail2 was suppressed by the specific knockdown of Numb PRR. In addition, the immunoprecipitation results showed that ATP11A could specifically bind to Numb PRR. The expression of ATP11A was also upregulated after TGFB stimulation, suggesting that the effect of ATP11A on EMT is TGFB dependent.
ATP11A is significantly upregulated in pancreatic cancer tissues, where it promotes the invasion and migration ability of pancreatic cancer cells. It is also associated with adverse prognosis in pancreatic cancer. Furthermore, ATP11A affects the epithelial-to-mesenchymal transition (EMT) of pancreatic cancer by regulating the TGFB dependent Numb PRR-ZEB1/Snail2 pathway.
NUMB 蛋白在肿瘤发生发展中起着至关重要的作用。本研究的主要目的是鉴定与胰腺癌生物学行为相关的 ATP11A，并阐明其与 NUMB 的关系及其内在机制。
首先，从癌症基因组图谱（TCGA）和基因-组织表达（GTE）数据库中获取的数据，用于研究胰腺癌细胞中 ATP11A mRNA 的表达及其与 NUMB mRNA 的关系。通过对 31 对胰腺癌组织和癌旁组织的 Western blot 检测，以及对 81 例胰腺癌标本的免疫组化检测，验证了 ATP11A 在胰腺癌组织中的蛋白水平表达。接下来，通过过表达或敲低 ATP11A 观察其对胰腺癌细胞侵袭和迁移能力的影响以及下游蛋白的变化。通过挽救实验确定了 ATP11A 影响 NUMB、ZEB1、Snail2 等蛋白的机制。此外，通过免疫沉淀实验探讨了 ATP11A 与 NUMB 之间的相互作用。最后，用 TGFB1 刺激胰腺癌细胞，检测 ATP11A 的表达，探讨 ATP11A 对 EMT 的影响是否依赖于 TGFB。
在 mRNA 水平上，胰腺癌组织中 ATP11A 的表达明显高于正常胰腺组织（<0.001）。ATP11A 的表达与 NUMB 的表达高度相关（R=0.676）。在蛋白水平上，胰腺癌组织中 ATP11A 的表达明显高于癌旁组织（=0.0009），且高 ATP11A 表达与预后不良相关。此外，我们的研究结果表明，ATP11A 可以促进胰腺癌细胞的侵袭和迁移。此外，ATP11A 可以正向调节 NUMB PRR、Snail2 和 ZEB1 蛋白的表达。挽救实验结果表明，通过 NUMB PRR 的特异性敲低，可以抑制 ATP11A 对 ZEB1/Snail2 的增强作用。此外，免疫沉淀实验结果表明，ATP11A 可以特异性结合 NUMB PRR。TGFB 刺激后，ATP11A 的表达也上调，提示 ATP11A 对 EMT 的影响依赖于 TGFB。
ATP11A 在胰腺癌组织中明显上调，促进了胰腺癌细胞的侵袭和迁移能力。它与胰腺癌的不良预后也有关。此外，ATP11A 通过调节 TGFB 依赖的 NUMB PRR-ZEB1/Snail2 通路影响胰腺癌细胞的上皮-间充质转化（EMT）。
